Pattern of eGFP expression and CD8⁺ T-cell immunity in vector-treated animals. (A) Mice treated with rAAVeGFP (top panel) and rAdeGFP (bottom panel) were analyzed for eGFP protein content of transduced muscle and frequency of eGFP_200-208 specific CD8⁺ T cells in spleen by IFN-γ ELISpot and tetramer analysis. Groups of 5 to 8 individual mice (with ± SEM) is shown (note differences in scale). eGFP gene expression in quadriceps muscle 252 days after vector administration was evaluated by direct imaging (B) and protein quantification by ELISA (C). Asterisk indicates that eGFP was below the ELISA detection limit. (D) Flow cytometric analysis of splenocytes from mice that were untreated (left) or inoculated intramuscularly 252 days earlier with rAAVeGFP (center) and rAdeGFP (right). Numbers represent the percentage of CD8⁺ T cells that stain with the H-2K^d eGFP_200-208 tetrameric complex. Gated on FSC/SSC appropriate for lymphocytes, propidium iodide–negative (PI⁻)CD4⁻B220⁻F4/80⁻CD3⁺ cells.